Cloning and Expression of Helicobacter pylori HpaA Gene
Objective: Helicobacter pylori is associated with chronic gastritis, peptic ulcers, gastric adenocarcinomaand gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Antibiotictherapies do not protect from potential re-infection and have a risk for development of drugresistance. Therefore, prophy...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Royan Institute (ACECR), Tehran
2009-01-01
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| Series: | Cell Journal |
| Subjects: | |
| Online Access: | http://celljournal.org/library/upload/article/Pirayeh.pdf |
| _version_ | 1828429070684127232 |
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| author | Moein Farshchian Saman Hoseinkhani Javad Atoofi Shahin Najar Peerayeh |
| author_facet | Moein Farshchian Saman Hoseinkhani Javad Atoofi Shahin Najar Peerayeh |
| author_sort | Moein Farshchian |
| collection | DOAJ |
| description | Objective: Helicobacter pylori is associated with chronic gastritis, peptic ulcers, gastric adenocarcinomaand gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Antibiotictherapies do not protect from potential re-infection and have a risk for development of drugresistance. Therefore, prophylactic vaccine mediated protection against H. pylori is an attractiveclinical interest. H. pylori adhesin A (HpaA) is a conserved surface lipoprotein and playsimportant roles in the pathogenesis of infection. In this study the recombinant protein (rHpaA)was over-expressed in E.coli.Materials and Methods: The hpaA gene was amplified by PCR. Prokaryote expression vectorpET28a-hpaA was constructed, and used to transform E.coli BL21DE3. The expressionof recombinant protein induced by IPTG was examined by SDS-PAGE. Western blot wereused to determine immunoreactivity of rHpaA by a rabbit polyclonal antibodies against wholecell of H. pylori.Results: The hpaA gene nucleotide sequence in the recombinant plasmid vector of pET-28-a-hpaA was consistent with that of H.pylori hpaA as published in the GenBank. SDS-PAGEdemonstrated that the constructed prokaryotic expression efficiently produced rHpaA at the1.5 mmol/L of IPTG. HpaA fusion protein was able to react with the rabbit polyclonal antibodyagainst whole cells of H. pylori.Conclusion: A prokaryotic expression system pET-28a-hpaA-BL21 with high efficiency of H.pylori hpaA gene was successfully established and the HpaA fusion protein showed satisfactoryimmunoreactivity. These results indicate that production of a specific recombinant proteinis an alternative and potentially more expeditious strategy for development of H. pylori vaccine. |
| first_indexed | 2024-12-10T17:27:37Z |
| format | Article |
| id | doaj.art-6c79c3dd2e2a4deb8e4af762bb752f3e |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-12-10T17:27:37Z |
| publishDate | 2009-01-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-6c79c3dd2e2a4deb8e4af762bb752f3e2022-12-22T01:39:46ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142009-01-01113273276Cloning and Expression of Helicobacter pylori HpaA GeneMoein FarshchianSaman HoseinkhaniJavad AtoofiShahin Najar PeerayehObjective: Helicobacter pylori is associated with chronic gastritis, peptic ulcers, gastric adenocarcinomaand gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Antibiotictherapies do not protect from potential re-infection and have a risk for development of drugresistance. Therefore, prophylactic vaccine mediated protection against H. pylori is an attractiveclinical interest. H. pylori adhesin A (HpaA) is a conserved surface lipoprotein and playsimportant roles in the pathogenesis of infection. In this study the recombinant protein (rHpaA)was over-expressed in E.coli.Materials and Methods: The hpaA gene was amplified by PCR. Prokaryote expression vectorpET28a-hpaA was constructed, and used to transform E.coli BL21DE3. The expressionof recombinant protein induced by IPTG was examined by SDS-PAGE. Western blot wereused to determine immunoreactivity of rHpaA by a rabbit polyclonal antibodies against wholecell of H. pylori.Results: The hpaA gene nucleotide sequence in the recombinant plasmid vector of pET-28-a-hpaA was consistent with that of H.pylori hpaA as published in the GenBank. SDS-PAGEdemonstrated that the constructed prokaryotic expression efficiently produced rHpaA at the1.5 mmol/L of IPTG. HpaA fusion protein was able to react with the rabbit polyclonal antibodyagainst whole cells of H. pylori.Conclusion: A prokaryotic expression system pET-28a-hpaA-BL21 with high efficiency of H.pylori hpaA gene was successfully established and the HpaA fusion protein showed satisfactoryimmunoreactivity. These results indicate that production of a specific recombinant proteinis an alternative and potentially more expeditious strategy for development of H. pylori vaccine.http://celljournal.org/library/upload/article/Pirayeh.pdfHelicobacter pylorihpaARecombinant ProteinpET28a |
| spellingShingle | Moein Farshchian Saman Hoseinkhani Javad Atoofi Shahin Najar Peerayeh Cloning and Expression of Helicobacter pylori HpaA Gene Cell Journal Helicobacter pylori hpaA Recombinant Protein pET28a |
| title | Cloning and Expression of Helicobacter pylori HpaA Gene |
| title_full | Cloning and Expression of Helicobacter pylori HpaA Gene |
| title_fullStr | Cloning and Expression of Helicobacter pylori HpaA Gene |
| title_full_unstemmed | Cloning and Expression of Helicobacter pylori HpaA Gene |
| title_short | Cloning and Expression of Helicobacter pylori HpaA Gene |
| title_sort | cloning and expression of helicobacter pylori hpaa gene |
| topic | Helicobacter pylori hpaA Recombinant Protein pET28a |
| url | http://celljournal.org/library/upload/article/Pirayeh.pdf |
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